Agent for inhibition of cytokine production and agent for inhibition of cell adhesion

ABSTRACT

The present invention provides an agent for inhibiting cytokine production or cell adhesion, comprising at least one compound selected from the group consisting of thiazole derivatives represented by the following general formula:  
                 
 
[wherein R 1  is a phenyl group which may have a lower alkoxy group(s) as a substituent(s) on the phenyl ring, and R 2  is a group represented by the following general formula:  
                 
 
(wherein R 3 &#39;s, which may be the same or different, are each a carboxyl group, a lower alkoxy group or the like) or the like] and salts thereof.

TECHNICAL FIELD

The present invention relates to an agent for inhibition of cytokineproduction and an agent for inhibition of cell adhesion.

BACKGROUND ART

A number of cytokines were discovered as protein factors which inhibitthe expression of human physiological activities such as immuneresponse, inflammation, hematopoiesis and the like, and their structuresand functions have gradually been made clear. As a result, it is beingclarified that the cytokines affect not only human immunological systembut also various other human physiological activities and further have aclose connection with the development, differentiation, homeostatis anddiseases of human body.

Many cytokines such as TNF-α, IL-1β, IL-6, IFN-γ and the like areidentified. It is known that they also have various pharmacologicalactivities.

Of the above cytokines, TNF-α (Tumor necrosis factor-α) was discoveredas an antineoplastic cytokine and was expected to be used as an.anticancer agent. However, TNF-α was later found to be the samesubstance as cachectin (a cachexia inducer) and is reported to have, forexample, a stimulating activity for production of IL-1 and othercytokines, an activity of proliferation of fibroblast, an endotoxinshock-inducing activity, an activity for increasing ICAM-1, ICAM-2(intercellular adhesion molecules), ELAM (endothelial leukocyte adhesionmolecule-1), etc. (these molecules are proteins for adhering leukocytesto endothelial cells) to accelerate the adhesion of leukocytes toendothelial cells, and an arthritis-causing activity such as boneresorption, cartilage destruction or the like [Beutler, B., et al.,Nature, 316, 552-554 (1985); Peetre, C., et al., J. Clin. Invest., 78,1694-1700 (1986); Kurt-Jones, E. A., et al., J. Immunol., 139, 2317-2324(1987); Bevilacqua, M. P., et al., Science, 241, 1160-1165 (1989);Akatu, K. & Suda, T., Medical Practice, 8 (9) 1393-1396 (1991)].

It is also reported that the concentration of TNF in blood or neurolymphincreases in infectious diseases by bacteria or parasites [Mitsuyama,M., Journal of Clinical and Experimental Medicine (IGAKU NO AYUMI), 159(8) 467-470 (1991); Nakao, M., Journal of Clinical and ExperimentalMedicine (IGAKU NO-AYUMI), 159 (8) 471-474 (1991)].

It is also reported that the activity of TNF is found in synovial fluidor serum, in chronic rheumatoid arthritis and that the activity is aTNF-α activity [Saxne, T., et al., Arthritis Rheum., 31, 1041 (1988);Chu, C. Q., et al., Arthritis Rheum., 34, 1125-1132 (1991); Macnaul, K.L., et al., J. Immunol., 145, 4154-4166 (1990); Brennan, F. M., et al.,J. Immunol., 22, 1907-1912 (1992); Brennan, F. M., et al., Bri. J.Rheum., 31, 293-298 (1992)].

It is also reported that the concentration of TNF is high in the sputaof patients of ARDS (acute respiratory distress syndrome) which is aserious respiratory disease [Millar, A. B., et al., Nature, 324, 73(1986)] and that TNF is associated with viral fulminant hepatitis [Muto,Y., et al., Lancet, ii, 72-74 (1986)].

It is also reported that the concentration of TNF-α in blood is high inmyocardial ischemia such as acute myocardial infarction [Latini, R., etal., J. Cardiovasc. Pharmacol., 23, 1-6 (1994)]. It is suggested thatTNF-α is associated with such a disease [Lefer, A. M., et al., Science,249, 61-64 (1990)]. It has recently been reported that TNF-α suppressesmyocardial contractility [Finkel, M. S., et al., Science, 257, 387-389(1992); Pagani, D. F., et al., J. Clin. Invest., 90, 389-398 (1992)].

Currently, no satisfactory chemotherapy is developed yet for theabove-mentioned various diseases such as chronic rheumatoid arthritis,endotoxin shock, ARDS and the like. To these diseases are merelyapplied, in a symptomatic treatment, steroidal agents, anti-inflammatoryagents, agents for inhibition of platelet agglutination, antibiotics,etc. As it was suggested as mentioned above that there is a closeconnection between the above diseases and the rise in concentration oractivity of TNF-α, it has recently been tried to apply TNF-α antibody orthe like to the diseases; however, such an approach has given nosatisfactory result, either. Therefore, it is desired in the art todevelop a drug for treatment of the above diseases, which can suppressthe excessive production of, in particular, TNF-α, according to a novelmechanism.

B cells are activated by antigen, proliferated and differentiated intoantibody-producing cells. IL-6 is known to be a cytokine participatingin this differentiation.

It is clear that IL-6 not only plays an important role in antibodyproduction of B cells, but also induces the proliferation anddifferentiation of T cells. It is also clear that IL-6 acts on livercells to induce the synthesis of proteins in acute phase, acts onhemopoietic cells to promote the formation of pluripotential colonies,and is an important factor in biophylactic systems such as immunesystem, hemopoietic system, nerve system, liver and the like.

As the diseases with which IL-6 is associated, there are mentioned aseries of autoimmune diseases such as hyper-γ-globulinemia, chronicrheumatoid arthritis, systemic lupus erythematosus (SLE) and the like;monoclonal B cell abnormal disease (e.g. myeloma); polyclonal B cellabnormal disease; atrial myxoma; Castleman syndrome; primaryglomerulonephritis; mesangial proliferative nephritis; cancerouscachexia; Lennander's lymphoma; psoriasis; Kaposi's sarcoma appearing inAIDS; postmenopausal osteoporosis; and so forth.

IL-1β is known to have various physiological activities. Specificexamples of these activities are inhibition of tumor cell, increase ofcytokine production from activated T cells, proliferation of fibroblast,synoviocyte and vessel endothelium, catabolism and thermacogenesis ofcell, differentiation of activated B cell, increase of NK activity,adhesion of neutrophils, anti-inflammation, inhibition of radiationdisorder, etc.

When IL-1β is produced at an increased rate and becomes excessive, IL-1βis thought to give rise to various diseases such as chronic rheumatoidarthritis, chronic inflammatory diseases and the like.

IFN is known to have various physiological activities and is actuallydetected in tissues and blood during many diseases. The diseases whoseonset is considered to have a close connection with IFN, includeviral-infectious diseases; infectious diseases by microorganisms otherthan viruses, chronic rheumatoid arthritis, collagen diseases (e.g.SLE), I-type allergy, uveitis, Behcet's disease, sarcoidosis,arteriosclerosis, diabetes, fulminant hepatitis, malignant tumor,Kawasaki disease, wounds of skin or mucosa, etc. [Journal of Clinicaland Experimental Medicine (IGAKU NO AYUMI), 174 (14), p. 1077, 1995].

Neutrophils express a bactericidal action to the enemy incoming intohuman body, by migration, phagocytosis, production of reactive oxygenand release of lysosomal enzymes. However, neutrophils are known toadhere to vascular endothelial cells and further infiltrate into tissuesduring the ischemia or reperfusion, or acute inflammation of varioustissues, leading to tissue disorder.

As stated above, various cytokines are known to cause various diseaseswhen the cytokines become excessive owing to, for example, theabnormally high production thereof. Therefore, it is desired toameliorate the abnormal state of cytokine to prevent or treat variousdiseases.

It is also desired to develop an agent for inhibiting the tissuedisorder caused by adhesion of neutrophils to vascular endothelialcells.

Some of the thiazole derivatives represented by the following generalformula (1):

(wherein R¹ is a phenyl group which may have a lower alkoxy group(s) asa substituent(s) on the phenyl ring; and R² is a group represented bythe following general formula:

[wherein R³'s, which may be the same or different, are each a carboxylgroup, a lower alkoxy group, a lower alkyl group, a lower alkenyl group,a group represented by -(A)_(l)-NR⁴R⁵ (A is a lower alkylene group; R⁴and R⁵, which may be the same or different, are each a hydrogen atom ora lower alkyl group; and l is 0 or 1), a hydroxyl group-substitutedlower alkyl group, a lower alkoxy group-substituted lower alkoxy group,a lower alkoxy group-substituted lower alkoxycarbonyl group or acarboxyl group-substituted lower alkoxy group; and m is an integer of1-3], or a heterocyclic ring residue having 1-2 hetero atoms selectedfrom the group consisting of nitrogen atom, oxygen atom and sulfur atom,which heterocyclic ring residue may have, as a substituent(s) on theheterocyclic ring, 1-3 groups selected from the group consisting ofcarboxyl group and lower alkoxy group) and salts thereof, are known in,for example, JP-A-5-51318 and JP-A-6-65222. These thiazole derivativesand salts thereof are also well-known to be useful as a reactive oxygeninhibitor.

DISCLOSURE OF THE INVENTION

The object of the present invention is to provide an agent forinhibiting the abnormally high production of cytokines or adhesion ofneutrophils to vascular endothelial cells, which satisfies therequirements of the art, i.e. an agent for inhibiting cytokineproduction or an agent for inhibiting cell adhesion.

The present inventor made a further study on the pharmacological actionsof the thiazole derivatives represented by the above general formula (1)and salts thereof. As a result, the present inventor found out thatthese thiazole derivatives and salts thereof can act as an agent forinhibiting cytokine production or an agent for inhibiting cell adhesion,both satisfying the above object of the present invention. The presentinvention has been completed based on the finding.

According to the present invention, there is provided an agent forinhibiting cytokine production, comprising, as the active ingredient, atleast one compound selected from the group consisting of thiazolederivatives represented by the above general formula (1) and saltsthereof.

According to the present invention, there is also provided an agent forinhibiting cell adhesion, comprising, as the active ingredient, at leastone compound selected from the group consisting of thiazole derivativesrepresented by the above general formula (1) and salts thereof.

According to the present invention, there is also provided an agent forinhibiting TNF-α production, comprising, as the active ingredient, atleast one compound selected from the group consisting of thiazolederivatives represented by the above general formula (1) and saltsthereof.

Of the thiazole derivatives represented by the general formula (1),preferred is6-[2-(3,4-diethoxyphenyl)thiazole-4-yl]pyridine-2-carboxylic acid.

As mentioned previously, some of the thiazole derivatives of the generalformula (1) and salts thereof and production processes thereof aredescribed in JP-A-5-51318 and JP-A-6-65222, and these thiazolederivatives are known to be useful as an agent for inhibiting reactiveoxygen. Meanwhile, the inhibition of cytokine production or celladhesion according to the present invention has no connection with theabove-mentioned inhibition of reactive oxygen by thiazole derivativesand is unpredictable from the inhibition of reactive oxygen.

The agent for inhibiting cytokine production or cell adhesion accordingto the present invention is useful for various diseases associated withthe abnormally high production of cytokines, particularly TNF-α, IL-1β,IL-6 and IFN-γ, or with increased adhesion. The present agent can besuitably used as a preventive or therapeutic agent particularly forchronic rheumatoid arthritis; endotoxin shock; ARDS caused by aspirationof gastric contents, toxic gas, sepsis, etc.; burn; asthma; myocardialinfarction in myocardial ischemia; viral myocarditis in acute phase;chronic heart failure (e.g. idiopathetic dilated cardiomyopathy); etc.The present agent can also be suitably used as a preventive ortherapeutic agent for ischemia-reperfusion injury caused at the time ofcoronary arterial bypass graft (CABG) or the use of artificial heartlung apparatus; shift from systemic inflammatory response syndrome(SIRS) toward organ failure (e.g. severe acute pancreatitis,disseminated intravasocular coagulation (DIC)); multiple organ failurecaused by hepatic insufficiency after hepatectomy such as resection ofhepatic cancer, or acute pancreatitis; severe acute pancreatitis;inflammatory bowel diseases such as ulcerative colitis, Crohn diseaseand the like; a series of autoimmune diseases such ashyper-γ-globulinemia, chronic rheumatoid arthritis, systemic lupuserythematosus (SLE), multiple sclerosis and the like; metastasis ofcancer; rejection in transplantation; monoclonal B cell abnormal disease(e.g. myeloma); polyclonal B cell abnormal disease; atrial myxoma;Castleman syndrome; primary glomerulonephritis; mesangial proliferativeglomerulonephritis; cancerous cachexia; Lennander's lymphoma; psoriasis;atopic dermatitis; Kaposi's sarcoma appearing in AIDS; postmenopausalosteoporosity; diabetes; sepsis; arteriosclerosis; and inflammatorydiseases (e.g. angitis and hepatitis).

Listed below are literatures relating to the diseases for which thepresent agent for inhibition of cytokine production or for inhibition ofcell adhesion is efficacious.

(1) Literatures Relating to Transplantation

-   -   (a) Kojima, Y. et al., (1993) Cardiovasc. Surg., 1, 577-582    -   (b) Yamataka, T. et al., (1993) J. Pediatr. Surg., 28, 1451-1457    -   (c) Stepkowshi, S. M. et al., (1994) J. Immunol., 153, 5336-5346        (2) Literatures Relating to Asthma    -   (a) Ohkawara, Y. et al., (1995) Am. J. Respir. Cell Mol. Biol.,        12, 4-12    -   (b) Chihara, J. et al., (1995) Immunol. Lett., 46, 241-244    -   (c) Hakansson, L. et al., (1995) J. Allergy Clin. Immunol., 96,        941-950        (3) Literatures Relating to Arteriosclerosis    -   (a) Poston, R. N. et al., (1992) Am. J. Pathol., 140, 665-673    -   (b) Ross, P., (1993) Nature, 362, 801-809    -   (c) Li, H. et al., (1993) Arterioscler. & Thromb., 13, 197-204    -   (d) Walpola, P. L. et al., (1995) Arterioscler. Thromb. Vasc.        Biol., 15, 2-10        (4) Literatures Relating to Metastasis of Cancer    -   (a) Garofalo, A. et al., (1995) Cancer Res., 55, 414-419    -   (b) Gardner, M. J. et al., (1995) Cancer Lett., 91, 229-234        (5) Literatures Relating to Diabetes    -   (a) McLeod, D. S. et al., (1995) Am. J. Pathol., 147, 642-653    -   (b) Schmidt, A. M. et al., (1995) J. Clin. Invest., 96,        1395-1403    -   (c) Jakubowski, A. et al., (1995) J. Immunol., 155, 938-946        (6) Literatures Relating to Multiple Sclerosis    -   (a) Dore-Duffy, P. et al., (1993) Adv. Exp. Med. Biol., 331,        243-248    -   (b) Mizobuchi, M. and Iwasaki, Y., (1994) Nippon Rinsho, 52,        2830-2836    -   (c) Cannella, B. and Raine, C. S., (1995) Ann. Neurol., 37,        424-435        (7) Literatures Relating to Multiple Organ Failure    -   (a) Law, M. M. et al., (1994) J. Trauma., 37, 100-109    -   (b) Anderson, J. A. et al., (1996) J. Clin. Invest., 97,        1952-1959        (8) Literatures Relating to Atopic Dermatitis    -   (a) Meng, H. et al., (1995) J. Cell Physiol., 165, 40-53    -   (b) Santamaria, L. F. et al., (1995) Int. Arch. Allergy        Immunol., 107, 359-362    -   (c) Wakita, H. et al., (1994) J. Cutan. Pathol., 21, 33-39        (9) Literatures Relating to Psoriasis    -   (a) Groves, R. W. et al., (1993) J. Am. Acad. Dermatol., 29,        67-72    -   (b) Uyemura K., (1993) J. Invest. Dermatol., 101, 701-705    -   (c) Lee, M. L. et al., (1994) Australas J. Dermatol., 35, 65-70    -   (d) Wakita, H. and Takigawa, M., (1994) Arch. Dermatol., 130,        457-463        (10) Literatures Relating to Chronic Rheumatoid Arthritis    -   (a) Hale, P. L. et al., (1993) Arthritis Rheum., 32, 22-30    -   (b) Iigo Y. et al., (1991) J. Immunol., 147, 4167-4171        (11) Literatures Relating to Acute Respiratory Distress Syndrome    -   (a) Tate, R. M. and Repine, J. E., (1983) Am. Rev. Respir. Dis.,        128, 552-559    -   (b) Beutler, B., Milsark, I. W. and Cerami, A. C., (1985)        Science, 229, 869-871    -   (c) Holman, R. G. and Maier, R. V., (1988) Arch. Surg., 123,        1491-1495    -   (d) Windsor, A. et al., (1993) J. Clin. Invest., 91, 1459-1468    -   (e) van der Poll, T. and Lowry, S. F., (1995) Prog. Surg. Basel.        Karger, 20, 18-32        (12) Literatures Relating to Ischemic Reperfusion Injury    -   (a) Yamazaki, T. et al., (1993) Am. J. Pathol., 143, 410-418    -   (b) Vaage, J. and Valen, G., (1993) Acand. J. Thorac.        Cardiovasc. Surg. Suppl., 41    -   (c) McMillen, M. A. et al., (1993) Am. J. Surg., 166, 557-562    -   (d) Bevilacqua, M. P. et al., (1994) Annu. Rev. Med., 45,        361-378    -   (e) Panes, J. and Granger, D. N., (1994) Dig. Dis., 12, 232-241        (13) Literatures Relating to Inflammatory Bowel Disease    -   (a), Mahida, Y. R. et al., (1989) Gut, 30, 835-838    -   (b) Nakamura, S. et al., (1993) Lab. Invest., 69, 77-85    -   (c) Beil, W. J. et al., (1995) J. Leukocyte Bio., 58, 284-298    -   (d) Jones, S. C. et al., (1995) Gut, 36, 724-730        (14) Literatures Relating to Systemic Inflammatory Response        Syndrome    -   (a) K. Mori and M. Ogawa, (1996) Molecular Medicine, 33, 9,        1080-1088    -   (b) Dinarello, C. A. et al., (1993) JAMA, 269, 1829

Specific examples of each of the groups used in the general formula (1)are as follows.

The phenyl group which may have a lower alkoxy group(s) as asubstituent(s) on the phenyl ring, include phenyl groups which may have1-3 straight chain or branched chain alkoxy groups of 1-6 carbon atomsas a substituent(s) on the phenyl ring, such as phenyl, 2-methoxyphenyl,3-methoxyphenyl, 4-methoxyphenyl, 2-ethoxyphenyl, 3-ethoxyphenyl,4-ethoxyphenyl, 4-isopropoxyphenyl, 4-pentyloxyphenyl, 4-hexyloxyphenyl,3,4-dimethoxyphenyl, 3-ethoxy-4-methoxyphenyl, 2,3-dimethoxyphenyl,3,4-diethoxyphenyl, 2,5-dimethoxyphenyl, 2,6-dimethoxyphenyl,3-propoxy-4-methoxyphenyl, 3,5-dimethoxyphenyl, 3,4-dipentyloxyphenyl,3,4,5-trimethoxyphenyl, 3-methoxy-4-ethoxyphenyl and the like.

The lower alkyl group includes straight chain or branched chain alkylgroups of 1-6 carbon atoms, such as methyl, ethyl, propyl, isopropyl,butyl, tert-butyl, pentyl, hexyl and the like.

The lower alkoxy group includes straight chain or branched chain alkoxygroups of 1-6 carbon atoms, such as methoxy, ethoxy, propoxy,isopropoxy, butoxy, tert-butoxy, pentyloxy, hexyloxy and the like.

The lower alkenyl group includes straight chain or branched chainalkenyl groups of 2-6 carbon atoms, such as vinyl, allyl, 2-butenyl,3-butenyl, 1-methylallyl, 2-pentenyl, 2-hexenyl and the like.

The group represented by -(A)_(l)-NR⁴R⁵ (A is a lower alkylene group; R⁴and R⁵, which may be the same or different, are each a hydrogen atom ora lower alkyl group; and l is 0 or 1) includes groups represented by-(A)_(l)-NR⁴R⁵ (A is an alkylene group of 1-6 carbon atoms; R⁴ and R⁵,which may be the same or different, are each a hydrogen atom or astraight chain or branched chain alkyl group of 1-6 carbon atoms; and lis 0 or 1), such as amino, methylamino, ethylamino, propylamino,isopropylamino, tert-butylamino, butylamino, pentylamino, hexylamino,dimethylamino, diethylamino, methylethylamino, methylpropylamino,aminomethyl, 2-aminoethyl, 3-aminopropyl, 4-aminobutyl, 5-aminopentyl,6-aminohexyl, 1,1-dimethyl-2-aminoethyl, 2-methyl-3-aminopropyl,methylaminomethyl, ethylaminomethyl, propylaminomethyl,butylaminomethyl, pentylaminomethyl, hexylaminomethyl,dimethylaminomethyl, 2-dimethylaminoethyl and the like.

The hydroxyl group-substituted lower alkyl group includes straight chainor branched chain alkyl groups of 1-6 carbon atoms having 1-3 hydroxylgroups, such as hydroxymethyl, 2-hydroxyethyl, 1-hydroxyethyl,1,2-dihydroxyethyl, 3-hydroxypropyl, 2,3-dihydroxypropyl,4-hydroxybutyl, 1,1-dimethyl-2-hydroxyethyl, 5,5,4-trihydroxypentyl,5-hydroxypentyl, 6-hydroxyhexyl, 1-hydroxyisopropyl,2-methyl-3-hydroxypropyl and the like.

The lower alkoxy group-substituted lower alkoxy group includesalkoxyalkoxy groups whose alkoxy moities are each a straight chain orbranched chain alkoxy group of 1-6 carbon atoms, such as methoxymethoxy,3-methoxypropoxy, ethoxymethoxy, 4-ethoxybutoxy, 6-propoxyhexyloxy,5-isopropoxypentyloxy, 1,1-dimethyl-2-butoxyethoxy,2-methyl-3-tert-butoxypropoxy, 2-pentyloxyethoxy, hexyloxymethoxy andthe like.

The lower alkoxycarbonyl group can be exemplified by straight chain orbranched chain alkoxycarbonyl groups of 1-6 carbon atoms, such asmethoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, isopropoxycarbonyl,butoxycarbonyl, tert-butoxycarbonyl, pentyloxycarbonyl, hexyloxycarbonyland the like.

The lower alkoxy group-substituted lower alkoxycarbonyl group includesalkoxy group-substituted alkoxycarbonyl groups whose alkoxy moities areeach a straight chain or branched chain alkoxy group of 1-6 carbonatoms, such as methoxymethoxycarbonyl, 3-methoxypropoxycarbonyl,ethoxymethoxycarbonyl, 4-ethoxybutoxycarbonyl,6-propoxyhexyloxycarbonyl, 5-isopropoxypentyloxycarbonyl,1,1-dimethyl-2-butoxy-ethoxycarbonyl,2-methyl-3-tert-butoxypropoxycarbonyl, 2-pentyloxyethoxycarbonyl,hexyloxymethoxycarbonyl and the like.

The carboxyl group-substituted lower alkoxy group includes carboxylgroup-substituted alkoxy groups whose alkoxy moiety is a straight chainor branched chain alkoxy group of 1-6 carbon atoms, such ascarboxymethoxy, 2-carboxyethoxy, 1-carboxyethoxy, 3-carboxypropoxy,4-carboxybutoxy, 5-carboxypentyloxy, 6-carboxyhexyloxy,1,1-dimethyl-2-carboxyethoxy, 2-methyl-3-carboxypropoxy and the like.

The heterocyclic ring residue having 1-2 hetero atoms selected from thegroup consisting of nitrogen atom, oxygen atom and sulfur atom includes,for example, pyrrolidinyl, piperidinyl, piperazinyl, morpholino,pyridyl, 1,2,5,6-tetrahydropyridyl, thienyl, quinolyl,1,4-dihydroquinolyl, benzothiazolyl, pyrazinyl, pyrimidyl, pyridazinyl,pyrrolyl, carbostyril, 3,4-dihydrocarbostyril,1,2,3,4-tetrahydroquinolyl, indolyl, isoindolyl, indolinyl,benzimidazolyl, benzoxazolyl, imidazolidinyl, isoquinolyl, quinazolinyl,quinoxalinyl, cinnolinyl, phthaladinyl, carbazolyl, acridinyl,chromanyl, isoindolinyl, isochromanyl, pyrazolyl, imidazolyl,pyrazolidinyl, phenothiazinyl, benzofuryl, 2,3-dihydro[b]furyl,benzothienyl, phenoxathienyl, phenoxazinyl, 4H-chromenyl, 1H-indazolyl,phenazinyl, xanthenyl, thianthrenyl, 2-imidazolinyl, 2-pyrrolinyl,furyl, oxazolyl, isooxazolyl, thiazolyl, isothiazolyl, pyranyl,2-pyrazolinyl, quinuclidinyl, 1,4-benzoxazinyl,3,4-dihydro-2H-1,4-benzoxazinyl, 3,4-dihydro-2H-1,4-benzthiazinyl,1,4-benzthiazinyl, 1,2,3,4-tetrahydro-quinoxalinyl,1,3-dithia-2,4-dihydronaphthalenyl, phenanthridinyl and1,4-dithianaphthalenyl.

The heterocyclic ring residue having 1-2 hetero atoms selected from thegroup consisting of nitrogen atom, oxygen atom and sulfur atom, whichhas 1-3 groups selected from the group consisting of carboxyl group andlower alkoxy groups, include, for example, 4-carboxy-2-furyl,5-carboxy-2-furyl, 4-carboxy-2-pyridyl, 6-carboxy-2-pyridyl,4-methoxy-5-carboxy-2-thiophenyl, 4-carboxy-2-thiazolyl,2-carboxy-4-pyridyl and 4-carboxy-2-pyrimidyl.

Of the thiazole derivatives represented by the general formula (1),those compounds having basic group react easily with pharmacologicallyacceptable ordinary acids to form respective salts. Such acids can beexemplified by inorganic acids such as sulfuric acid, nitric acid,hydrochloric acid, phosphoric acid, hydrobromic acid and the like; andorganic acids such as acetic acid, p-toluenesulfonic acid,ethanesulfonic acid, oxalic acid, maleic acid, fumaric acid, malic acid,tartaric acid, citric acid, succinic acid, benzoic acid and the like.

Of the thiazole derivatives represented by the general formula (1),those compounds having acidic group react easily with pharmacologicallyacceptable ordinary basic compounds to form respective salts. Such basiccompounds include, for example, sodium hydroxide, potassium hydroxide,calcium hydroxide, sodium carbonate and potassium hydrogencarbonate.

Needless to say, the compounds of the present invention include opticalisomers.

Each of the compounds of the general formula (1) is used generally inthe form of ordinary pharmaceutical preparation. The pharmaceuticalpreparation is prepared by using diluents or excipients ordinarily used,such as filler, bulking agent, binder, humectant, disintegrator,surfactant, lubricant and the like. The pharmaceutical preparation canbe prepared in various forms depending upon the purpose of remedy, andthe typical forms include tablets, pills, a powder, a solution, asuspension, an emulsion, granules, capsules, suppositories, an injection(e.g. solution or suspension), etc. In preparing tablets, there can beused various carriers known in the art. The carriers can be exemplifiedby excipients such as lactose, white sugar, sodium chloride, glucose,urea, starch, calcium carbonate, kaolin, crystalline cellulose, silicicacid and the like; binders such as water, ethanol, propanol, simplesyrup, glucose solution, starch solution, gelatin solution,carboxymethyl cellulose, shellac, methyl cellulose, potassium phosphate,polyvinylpyrrolidone and the like; disintegrators such as dry starch,sodium alginate, powdered agar, powdered laminarin, sodiumhydrogencarbonate, calcium carbonate, polyoxyethylene sorbitan-fattyacid esters, sodium lauryl sulfate, stearic acid monoglyceride, starch,lactose and the like; disintegration inhibitors such as white sugar,stearin, cacao butter, hydrogenated oil and the like; absorptionpromoters such as quaternary ammonium salts, sodium lauryl sulfate andthe like; humectants such as glycerine, starch and the like; adsorbentssuch as starch, lactose, kaolin, bentonite, colloidal silicic acid andthe like; and lubricants such as refined talc, stearic acid salts, boricacid powder, polyethylene glycol and the like. The tablets can beprepared, as necessary, in the form of ordinary coated tablets, such assugar-coated tablets, gelatin-coated tablets, enteric coated tablets orfilm-coated tablets, or in the form of double-layered tablets ormulti-layered tablets. In preparing pills, there can be used variouscarriers known in the art. The carriers can be exemplified by excipientssuch as glucose, lactose, starch, cacao butter, hardened vegetable oils,kaolin, talc and the like; binders such as powdered acacia, powderedtragacanth, gelatin, ethanol and the like; and disintegrators such aslaminarin, agar and the like. In preparing suppositories, there can beused various carriers known in the art. The carriers can be exemplifiedby a polyethylene glycol, cacao butter, a higher alcohol, a higheralcohol ester, gelatin and a semi-synthetic glyceride. Capsules can beprepared ordinarily by mixing the above-mentioned active ingredient withvarious carriers mentioned above and filling the resulting mixture intohard gelatin capsules, soft capsules or the like, according to anordinary method. In preparing an injection (solution, emulsion orsuspension), it is sterilized and is preferably made isotonic to theblood. In preparing the solution, emulsion or suspension, there can beused all diluents ordinarily used in the art, such as water, ethylalcohol, macrogol, propylene glycol, ethoxylated isostearyl alcohol,polyoxy-isostearyl alcohol and polyoxyethylene sorbitan-fatty acidesters. In this case, the injection may contain sodium chloride, glucoseor glycerine in an amount sufficient to make the injection isotonic, andmay further contain a solubilizing adjuvant, a buffer solution, asoothing agent, etc. all ordinarily used. The pharmaceutical preparationmay furthermore contain, as necessary, a coloring agent, a preservative,a perfume, a flavoring agent, a sweetening agent and other drugs.

The amount of the active ingredient compound to be contained in thepharmaceutical preparation of the present invention is not particularlyrestricted and can be appropriately selected from a wide range, but thedesirable amount is generally about 1-70% by weight in thepharmaceutical preparation.

The method for administering the pharmaceutical preparation of thepresent invention is not particularly restricted. The method is decideddepending upon the form of preparation, the age, sex and otherconditions of patient, the disease condition of patient, etc. Forexample, tablets, pills, a solution, a suspension, an emulsion, granulesor capsules are administered orally. An injection is intravenouslyadministered singly or in admixture with an ordinary auxiliary solutionof glucose, amino acids or the like, or, as necessary, is singlyadministered intramuscularly, intradermally, subcutaneously orintraperitoneally. Suppositories are administered intrarectally.

The dose of the pharmaceutical preparation of the present invention isappropriately selected depending upon the administration method, theage, sex and other conditions of patient, the disease condition ofpatient, etc., but the desirable dose is generally about 0.2-200 mg perkg of body weight per day in terms of the amount of the activeingredient, i.e. the compound of general formula (1) or the saltthereof.

EXAMPLES

The present invention is hereinafter described specifically by way ofReference Examples, Examples, Pharmacological Tests and PreparationExamples.

Reference Example 1

To a solution of 0.88 g of 6-acetyl-3-acetyloxy-2-ethoxycarbonylpyridinein 8.8 ml of acetic acid was added 0.19 ml of bromine dropwise, and themixture was stirred at 75° C. for 5 minutes. Evaporation of the solventgave 0.77 g of 6-(2-bromoacetyl)-2-ethoxycarbonyl-3-hydroxypyridinehydrobromide.

Reference Example 2

5-(2-Bromoacetyl)-2-methoxycarbonylfuran was prepared from5-acetyl-2-methoxycarbonylfuran using the procedure given in ReferenceExample 1.

Reference Example 3

A solution of 29 g of 3,4-diethoxybenzonitrile and 23 g of thioacetamidein 120 ml of 10% hydrochloric acid-DMF was stirred at 90° C. for 3 hoursand then 130° C. for 5 hours. After evaporation of the solvent, theresidue was washed with diethyl ether (2×100 ml) and water (2×100 ml).The resulting crystals were collected by filtration and dried to obtain21.7 g of 3,4-diethoxythiobenzamide.

Reference Example 4

4-Methoxy-3-propoxythiobenzamide was prepared from4-methoxy-3-propoxybenzonitrile using the procedure given in ReferenceExample 3.

Reference Example 5

To a solution of 877 mg of 5-(2-bromoacetyl)-2-methoxycarbonylfuran in40 ml of methanol was added 800 mg of 4-methoxy-3-propoxythiobenzamide,and the mixture was refluxed for 1 hour. The reaction mixture wasconcentrated approximately 1/4, then added diethyl ether. After coolingthe solution, a precipitate was collected by filtration and dried toobtain 1.05 g of2-(4-methoxy-3-propoxyphenyl)-4-(5-methoxycarbonyl-2-furyl)thiazole as abrown powder. mp. 141.0-142.0° C.

Reference Examples 6-36

Using appropriate starting materials and using procedures similar tothose used in the above Reference Examples, there were obtained thecompounds shown in Table 1 to Table 6. TABLE 1

Reference Example R¹ R² Properties 6

Melting point: 141.0-142.0° C. Brown powder 7

Melting point: 138.0-139.0° C. Light yellow powder 8

Melting point: 82.5-85.0° C. Light yellow powder 9

Melting point: 83.5-85.5° C. White powder 10

Identical with the properties of a compound mentioned in JP-A-5-51318 11

Melting point: 95.0-97.5° C. White powder

TABLE 2 Reference Example R¹ R² Properties 12

Light brown oil NMR (1) 13

Light yellow viscous oil NMR (2) 14

Colorless viscous oil NMR (3) 15

Colorless viscous oil NMR (4) 16

Colorless viscous oil NMR (5) 17

Colorless viscous oil NMR (6)

TABLE 3 Reference Example R¹ R² Properties 18

Melting point: 104.0-106.5° C. Light yellow needles 19

Melting point: 158.5-159.5° C. Light yellow powder 20

Light brown solid NMR (7) 21

Light yellow oil NMR (8) 22

Melting point: 179.5-180.5° C. Light brown needles 23

Melting point: 165.0-167.0° C. White powder

TABLE 4 Reference Example R¹ R² Properties 24

Colorless amorphous NMR (9) 25

Yellow amorphous NMR (10) 26

Light yellow viscous oil NMR (11) 27

Light yellow powder NMR (12) 28

Light yellow viscous oil NMR (13)

TABLE 5 Reference Example R¹ R² Properties 29

Melting point: 106.0-107.0° C. Light yellow powder 30

Yellow oil NMR (14) 31

Colorless viscous oil NMR (15) 32

Colorless viscous oil NMR (16)

TABLE 6 Reference Example R¹ R² Properties 33

Brown oil NMR (17) 34

Light yellow oil NMR (18) 35

Melting point: 101.5-105.5° C. Light yellow powder

The above-obtained compounds had the following NMR spectra.

NMR(1): ¹H-NMR (CDCl₃) δ ppm; 1.49 (3H, t, J=7.0 Hz), 1.51 (3H, t, J=7.0Hz), 1.86 (3H, d, J=1.2 Hz), 1.98 (3H, d, J=1.2 Hz), 3.81 (3H, s), 3.95(3H, s), 4.12 (2H, q, J=7.0 Hz), 4.22 (2H, q, J=7.0 Hz), 6.36 (1H,br-s), 6.92 (1H, d, J=8.3 Hz), 7.37 (1H, s), 7.53 (1H, dd, J=2.0 Hz,J=8.3 Hz), 7.61 (1H, d, J=2.0 Hz), 7.97 (1H, d, J=2.3 Hz), 8.22 (1H, d,J=2.3 Hz).

NMR(2): ¹H-NMR (CDCl₃) δ ppm; 1.50 (3H, t, J=7.0 Hz), 1.52 (3H, t, J=7.0Hz), 1.74-2.04 (3H, m), 2.86 (2H, t, J=7.7 Hz), 3.58-3.72 (2H, m), 3.89(3H, s), 3.96 (3H, s), 4.16 (2H, q, J=7.0 Hz), 4.23 (2H, q, J=7.0 Hz),6.93 (1H, d, J=8.4 Hz), 7.40 (1H, s), 7.54 (1H, dd, J=2.1 Hz, J=8.4 Hz),7.60 (1H, d, J=2.1 Hz), 8.02 (1H, d, J=2.3 Hz), 8.23 (1H, d, J=2.4 Hz).

NMR(3): ¹H-NMR (CDCl₃) δ ppm; 1.49 (3H, t, J=7.0 Hz), 1.52 (3H, t, J=7.0Hz), 3.53 (2H, d, J=6.4 Hz), 3.86 (3H, s), 3.96 (3H, s), 4.16 (2H, q,J=7.0 Hz), 4.23 (2H, q, J=7.0 Hz), 5.02-5.21 (2H, m), 5.91-6.19 (1H, m),6.93 (1H, d, J=8.4 Hz), 7.39 (1H, s), 7.53 (1H, dd, J=2.1 Hz, J=8.4 Hz),7.61 (1H, d, J=2.1 Hz), 7.98 (1H, d, J=2.4 Hz), 8.26 (1H, d, J=2.4 Hz).

NMR(4): ¹H-NMR (CDCl₃) δ ppm; 1.27 (6H, s), 1.50 (3H, t, J=7.0 Hz), 1.51(3H, t, J=7.0 Hz), 2.61 (1H, br-s), 2.95 (2H, s), 3.89 (3H, s), 3.96(3H, s), 4.16 (2H, q, J=7.0 Hz), 4.22 (2H, q, J=7.0 Hz), 6.93 (1H, d,J=8.3 Hz), 7.40 (1H, s), 7.54 (1H, dd, J=2.1 Hz), J=8.3 Hz), 7.59 (1H,d, J=2.1HZ), 8.00 (1H, d, J=2.4 Hz), 8.31 (1H, d, J=2.4 Hz).

NMR(5): ¹H-NMR (CDCl₃) δ ppm; 1.29 (3H, d, J=6.2 Hz), 1.49 (3H, t, J=7.0Hz), 1.52 (3H, t, J=7.0 Hz), 2.08 (1H, br-s), 2.75-3.05 (2H, m), 3.89(3H, s), 3.97 (3H, s), 4.08-4.29 (1H, m), 4.16 (2H, q, J=7.0 Hz), 4.23(2H, q, J=7.0 Hz), 6.93 (1H, d, J=8.4 Hz), 7.40 (1H, s), 7.54 (1H, dd,J=2.1 Hz, J=8.4 Hz), 7.61 (1H, d, J=2.1 Hz), 8.02 (1H, d, J=2.3 Hz),8.28 (1H, d, J=2.3 Hz).

NMR(6): ¹H-NMR (CDCl₃) δ ppm; 1.23 (3H, d, J=6.2 Hz), 1.49 (3H, t, J=7.0Hz), 1.51 (3H, t, J=7.0 Hz), 1.71-1.98 (3H, m), 2.86 (2H, t, J=8.0 Hz),3.69-3.86 (1H, m), 3.89 (3H, s), 3.96 (3H, s), 4.16 (2H, q, J=7.0 Hz),4.23 (2H, q, J=7.0 Hz), 6.93 (1H, d, J=8.4 Hz), 7.40 (1H, s), 7.54 (1H,dd, J=2.1 Hz, J=8.4 Hz), 7.60 (1H, d, J=2.1 Hz), 8.01 (1H, d, J=2.3 Hz),8.23 (1H, d, J=2.3 Hz).

NMR(7): ¹H-NMR (CDCl₃) δ ppm; 1.49 (3H, t, J=7.0 Hz), 1.51 (3H, t, J=7.0Hz), 2.30 (6H, s), 3.56 (2H, s), 3.88 (3H, s), 3.96 (3H, s), 4.16 (2H,q, J=7.0 Hz), 4.23 (2H, q, J=7.0 Hz), 6.92 (1H, d, J=8.4 Hz), 7.43 (1H,s), 7.54 (1H, dd, J=2.1 Hz, J=8.4 Hz), 7.61 (1H, d, J=2.1 Hz), 8.15 (1H,d, J=2.4 Hz), 8.35 (1H, d, J=2.4 Hz).

NMR(8): ¹H-NMR (CDCl₃) δ ppm; 1.49 (3H, t, J=7.0 Hz), 1.51 (3H, t, J=7.0Hz), 3.90 (3H, s), 3.96 (3H, s), 3.99 (2H, s), 4.15 (2H, q, J=7.0 Hz),4.22 (2H, q, J=7.0 Hz), 6.92 (1H, d, J=8.3 Hz), 7.43 (1H, s), 7.53 (1H,dd, J=2.1 Hz, J=8.3 Hz), 7.59 (1H, d, J=2.1 Hz), 8.14 (1H, d, J=2.3 Hz),8.29 (1H, d, J=2.3 Hz).

NMR(9): ¹H-NMR (CDCl₃) δ ppm; 1.19 (3H, s), 1.50 (3H, t, J=7.0 Hz), 1.52(3H, t, J=7.0 Hz), 2.72 (1H, t, J=6.8 Hz), 2.91 (1H, d, J=13.5 Hz), 3.01(1H, s), 3.07 (1H, d, J=13.5 Hz), 3.37 (2H, dd, J=2.1 Hz, J=6.8 Hz),3.92 (1H, s), 3.97 (1H, s), 4.16 (2H, q, J=7.0 Hz), 4.22 (2H, q, J=7.0Hz), 6.93 (1H, d, J=8.3 Hz), 7.42 (1H, s), 7.54 (1H, dd, J=2.1 Hz, J=8.3Hz), 7.59 (1H, d, J=2.1 Hz), 8.02 (1H, d, J=2.3 Hz), 8.32 (1H, d, J=2.3Hz).

NMR(10): ¹H-NMR (CDCl₃) δ ppm; 1.50 (3H, t, J=7.0 Hz), 1.52 (3H, t,J=7.0 Hz), 1.58 (3H, d, J=6.5 Hz), 2.32 (1H, d, J=4.2 Hz), 3.91 (3H, s),3.97 (3H, s), 4.16 (2H, q, J=7.0 Hz), 4.22 (2H, q, J=7.0 Hz), 5.21-5.38(1H, m), 6.92 (1H, d, J=8.4 Hz), 7.43 (1H, s), 7.54 (1H, dd, J=2.1 Hz,J=8.4 Hz), 7.61 (1H, d, J=2.1 Hz), 8.25 (1H, d, J=2.2 Hz), 8.33 (1H, d,J=2.2 Hz).

NMR(11): ¹H-NMR (CDCl₃) δ ppm; 1.41-1.59 (9H, m), 1.94 (3H, dd, J=1.6Hz, J=6.6 Hz), 6.22-6.51 (1H, m), 6.68-6.85 (1H, m), 6.92 (1H, d, J=8.4Hz), 7.32 (1H, s), 7.41 (1H, d, J=2.0 Hz), 7.54 (1H, dd, J=2.0 Hz, J=8.4Hz), 7.60 (2H, d, J=2.0 Hz).

NMR(12): ¹H-NMR (CDCl₃) δ ppm; 1.49 (3H, t, J=7.0 Hz), 1.50 (3H, t,J=7.0 Hz), 1.51 (3H, t, J=7.0 Hz), 3.47 (2H, d, J=6.4 Hz), 3.87 (1H, s),4.16 (2H, q, J=7.0 Hz), 4.19 (2H, q, J=7.0 Hz), 4.22 (2H, q, J=7.0 Hz),5.00-5.19 (2H, m), 5.91-6.15 (1H, m), 6.92 (1H, d, J=8.4 Hz), 7.30 (1H,s), 7.33 (1H, d, J=2.0 Hz), 7.44 (1H, d, J=2.0 Hz), 7.53 (1H, dd, J=2.1Hz, J=8.4 Hz), 7.60 (1H, d, J=2.1 Hz).

NMR(13): ¹H-NMR (CDCl₃) δ ppm; 1.08 (3H, t, J=7.5 Hz), 1.50 (3H, t,J=7.0 Hz), 1.82-2.05 (5H, m), 3.85 (3H, s), 3.93 (3H, s), 4.11 (2H, t,J=6.9 Hz), 4.19 (2H, q, J=7.0 Hz), 6.22-6.51 (1H, m), 6.65-6.83 (1H, m),6.93 (1H, d, J=8.3 Hz), 7.33 (1H, s), 7.41 (1H, d, J=2.0 Hz), 7.55 (1H,dd, J=2.0 Hz, J=8.3 Hz), 7.60 (2H, d, J=2.0 Hz).

NMR(14): ¹H-NMR (CDCl₃) δ ppm; 1.49 (3H, t, J=7.0 Hz), 1.51 (3H, t,J=7.0 Hz), 1.78 (3H, s), 3.47 (2H, s), 3.85 (3H, s), 3.96 (3H, s), 4.16(2H, q, J=7.0 Hz), 4.23 (2H, q, J=7.0 Hz), 4.69 (1H, s), 4.88 (1H, s),6.92 (1H, d, J=8.4 Hz), 7.39 (1H, s), 7.53 (1H, dd, J=2.1 Hz, J=8.4 Hz),7.61 (1H, d, J=2.1 Hz), 7.96 (1H, d, J=2.3 Hz), 8.28 (1H, d, J=2.3 Hz).

NMR(15): ¹H-NMR (CDCl₃) δ ppm; 0.95 (6H, d, J=6.6 Hz), 1.49 (3H, t,J=7.0 Hz), 1.51 (3H, t, J=7.0 Hz), 1.90-2.14 (1H, m), 2.61 (2H, d, J=7.3Hz), 3.85 (3H, s), 3.96 (3H, S), 4.15 (2H, q, J=7.0 Hz), 4.22 (2H, q,J=7.0 Hz), 6.92 (1H, d, J=8.3HZ), 7.38 (1H, s), 7.55 (1H, dd, J=2.1 Hz,J=8.3 Hz), 7.60 (1H, d, J=2.1 Hz), 7.93 (1H, d, J=2.4 Hz), 8.23 (1H, d,J=2.4 Hz).

NMR(16): ¹H-NMR (CDCl₃) δ ppm; 1.01 (3H, t, J=7.4 Hz), 1.44 (3H, t,J=7.1 Hz), 1.49 (3H, t, J=7.0 Hz), 1.51 (3H, t, J=7.0 Hz), 1.71 (2H,sextet, J=7.4 Hz), 2.72 (2H, t, J=7.4 Hz), 3.87 (3H, s), 4.16 (2H, q,J=7.0 Hz), 4.22 (2H, q, J=7.0 Hz), 4.43 (2H, q, J=7.1 Hz), 6.92 (1H, d,J=8.4 Hz), 7.39 (1H, s), 7.53 (1H, dd, J=2.1 Hz, J=8.4 Hz), 7.62 (1H, d,J=2.1 Hz), 7.97 (1H, d, J=2.3 Hz), 8.21 (1H, d, J=2.3 Hz).

NMR(17): ¹H-NMR (CDCl₃) δ ppm; 1.49 (3H, t, J=7.0 Hz), 1.51 (3H, t,J=7.0 Hz), 1.97 (3H, dd, J=1.6 Hz, J=6.5 Hz), 3.85 (3H, s), 3.96 (3H,s), 4.16 (2H, q, J=7.0 Hz), 4.23 (2H, q, J=7.0 Hz), 6.41 (1H, dq, J=6.5Hz, J=15.9 Hz), 6.75 (1H, dd, J=1.6 Hz, J=15.9 Hz), 6.93 (1H, d, J=8.3Hz), 7.40 (1H, s), 7.55 (1H, dd, J=2.1 Hz, J=8.3 Hz), 7.60 (1H, d, J=2.1Hz), 8.21 (2H, s).

NMR(18): ¹H-NMR (CDCl₃) δ ppm; 1.49 (3H, t, J=7.0 Hz), 1.51 (3H, t,J=7.0 Hz), 3.87 (3H, s), 3.96 (3H, s), 4.16 (2H, q, J=7.0 Hz), 4.23 (2H,q, J=7.0 Hz), 5.44 (1H, dd, J=1.1 Hz, J=11.1 Hz), 5.92 (1H, dd, J=1.1Hz, J=17.7 Hz), 6.93 (1H, d, J=8.3 Hz), 7.09 (1H, dd, J=11.1 Hz, J=17.7Hz), 7.42 (1H, s), 7.54 (1H, dd, J=2.1 Hz, J=8.3 Hz), 7.61 (1H, d, J=2.1Hz), 8.28 (2H, br-s).

Example 1

To a suspension of 970 mg of2-(4-methoxy-3-propoxyphenyl)-4-(5-methoxycarbonyl-2-furyl)thiazole in30 ml of methanol was added 20 ml of 1,4-dioxane and 5 ml of a 5 Naqueous sodium hydroxide solution. The reaction mixture was refluxed for3 hours, then concentrated approximately 1/10. Water was added to theresidue, and washed with ethyl acetate. To the aqueous layer wasacidified with 5 N hydrochloric acid, and extracted with ethyl acetate.The combined organic layer was washed with water and a saturated aqueoussodium chloride solution, and dried with magnesium sulfate. Evaporationthe solution, the residue was recrystallized from ethyl acetate toobtain 420 mg of2-(4-methoxy-3-propoxyphenyl)-4-(5-carbonyl-2-furyl)thiazole as a whitepowder. mp. 191.0-192.0° C.

Examples 2-35

Using appropriate starting materials and using procedures similar tothat used in Example 1, there were obtained the compounds shown in Table7 to Table 12. TABLE 7

Example R¹ R² Properties 1

Melting point: 191.0-192.0° C. White powder 2

Melting point: 182.0-184.0° C. White powder 3

Melting point: 163.0-167.0° C. Light yellow powder 4

Melting point: 202.0-203.0° C. White powder 5

Melting point: 201.0-202.0° C. White powder 6

Melting point: 153.0-154.0° C. Light yellow granules

TABLE 8 Example R¹ R² Properties 7

Light yellow amorphous NMR (1) 8

Melting point: 109.5-111.5° C. White powder 9

Melting point: 137.0-139.0° C. White powder 10

Melting point: 135.0-138.0° C. White powder 11

Melting point: 110.0-112.5° C. White powder 12

Melting point: 165.0-167.0° C. Colorless needles

TABLE 9 Example R¹ R² Properties 13

Melting point: 204.5-206.5° C. Colorless needles 14

Form: monohydro- chloride Yellow amorphous NMR (2) 15

Melting point: 201.0-202.0° C. Light yellow needles 16

Melting point: 206.0-207.0° C. White powder 17

Melting point: 134.0-136.0° C. White powder 18

Melting point: 189.0-190.0° C. Light yellow plates

TABLE 10 Example R¹ R² Properties 19

Melting point: 147.5-149.0° C. Light yellow prisms 20

Melting point 139.0-141.0° C. Light yellow prisms 21

Identical with the properties of a compound mentioned in JP-A-5-51318 22

Identical with the properties of a compound mentioned in JP-A-5-51318 23

Identical with the properties of a compound mentioned in JP-A-5-51318 24

Identical with the properties of a compound mentioned in JP-A-5-51318 25

Identical with the properties of a compound mentioned in JP-A-6-65222

TABLE 11 Example R¹ R² Properties 26

Identical with the properties of a compound mentioned in JP-A-6-65222 27

Identical with the properties of a compound mentioned in JP-A-6-65222 28

Identical with the properties of a compound mentioned in JP-A-6-65222 29

Melting point: 65.0-68.0° C. Light yellow powder 30

Melting point: 163.0-165.0° C. White powder 31

Melting point: 145.0-147.0° C. White powder

TABLE 12 Example R¹ R² Properties 32

Melting point: 176.0-179.0° C. Colorless needles 33

Melting point: 208.0-210.0° C. White powder 34

Melting point: 175.5-177.5° C. Colorless prisms 35

Melting point: 188.5-190.0° C. White powder

The above-obtained compounds had the following NMR spectra.

NMR(1): ¹H-NMR (CDCl₃) δ ppm; 1.14 (3H, s), 1.35 (3H, s), 1.49 (3H, t,J=7.0 Hz), 1.50 (3H, t, J=7.0 Hz), 3.84 (3H, s), 4.15 (2H, q, J=7.0 Hz),4.21 (2H, q, J=7.0 Hz), 4.96 (1H, s), 6.91 (1H, d, J=8.3 Hz), 7.44 (1H,s), 7.52 (1H, dd, J=2.1 Hz, J=8.3 Hz), 7.58 (1H, d, J=2.1 Hz), 8.37 (1H,d, J=2.4 Hz), 8.55 (1H, d, J=2.4 Hz).

NMR(2): ¹H-NMR (DMSO-D₆) δ ppm; 1.34 (3H, t, J=6.8 Hz), 1.36 (3H, t,J=6.8 Hz), 2.74 (3H, s), 2.76 (3H, s), 3.86 (3H, s), 4.08 (2H, q, J=6.8Hz), 4.14 (2H, q, J=6.8 Hz), 4.29-4.56 (2H, m), 7.06 (1H, d, J=8.9 Hz),7.35-7.72 (2H, m), 8.16 (1H, s), 8.39 (1H, d, J=1.9 Hz), 8.67 (1H, d,J=1.9 Hz), 10.95 (1H, br-s).

Pharmacological Test 1 (Adhesion-Inhibiting Action 1)

A test compound was dissolved in 0.1 M sodium hydroxide. To theresulting solution was added a 9-fold volume of PBS (phosphate bufferedsaline) of Dulbecco formula (a product of Takara Co.) to prepare a 1 mMtest compound solution. This solution was diluted with 0.1 M sodiumhydroxide/PBS (1:9) to prepare a 0.1 mM test compound solution and a0.01 mM test compound solution. The two test compound solutions wereeach diluted 40-fold with a RPMI-1640 medium [containing 10% FCS (fetalcalf serum)]. Separately, N-formylmethionylleucylphenylalanine (fMLP) (2mM dissolved in dimethylformamide) was diluted with the RPMI-1640 medium(containing 10% FCS) to prepare a 0.25 mM fMLP solution.

Purified neutrophils were obtained from the whole blood of healthyperson by dextran sedimentation, Ficoll-Paque-density density gradientcentrifugation and erythrocycte hemolysis; then, were suspended in PBS(3 ml); and labelled with 50 μl of a fluorescence-labelling agent(BCECF-AM, a product of Dojindo Lab.) at room temperature for 1 hour.Human umbilical vein endothelial cells (HUVEC) (a product of CloneticsCo.) were cultivated on a 24-well culture plate, and a test was startedwhen the cells became confluent.

The medium in each well of the culture plate was removed. To the wellswere added 0.2 ml of RPMI-1640 (containing 10% FCS) or 0.2 ml of thediluted test compound solution, and 0.2 ml of the fMLP solution. Lastly,10⁶ fluorescence-labelled neutrophils were added to each well, and eachresulting mixture was incubated at 37° C. for 30 minutes. Adherentneutrophils and non-adherent neutrophils cells were collected separatelyand measured for fluorescent intensity. Using a separately preparedstandard line between number of neutrophils and fluorescent intensity,the number of cells was determined and a test compound concentration of50% adhesion inhibition, i.e. IC₅₀ was determined.

The results are shown in Table 13. TABLE 13 Test compound IC₅₀ (μM)Compound of Example 1 >10 Compound of Example 2 0.6 Compound of Example3 >10 Compound of Example 4 8.5 Compound of Example 5 <0.1 Compound ofExample 6 <0.1 Compound of Example 7 >10 Compound of Example 8 >10Compound of Example 9 >10 Compound of Example 10 >10 Compound of Example11 >10 Compound of Example 12 2.5 Compound of Example 13 3.0 Compound ofExample 21 >10 Compound of Example 22 >10 Compound of Example 23 5.6Compound of Example 24 <0.1 Compound of Example 25 5.0 Compound ofExample 27 2.9 Compound of Example 29 <0.1 Compound of Example 30 4.4Compound of Example 31 0.5 Compound of Example 32 0.1 Compound ofExample 33 0.95 Compound of Example 34 5.9 Compound of Example 35 0.8Pharmacological Test 2[Adhesion-Inhibiting Action 2 (Action on Appearance of ICAM-1 or VCAM-1to Endothelial Cells]

A test compound was dissolved in 0.1 M sodium hydroxide. To theresulting solution was added a 9-fold volume of PBS of Dulbecco formula(a product of Takara Co.) to prepare a 1 mM test compound solution. Thissolution was diluted with 0.1 M sodium hydroxide/PBS (1:9) to preparesolutions containing 300 μM, 100 μM, 30 μM, 10 μM and 3 μM of the testcompound, respectively. The solutions were each diluted 10-fold withRPMI-1640 (containing 10% FCS) to prepare 100 μM, 30 μM, 10 μM, 3 μM, 1μM and 0.3 μM of test compound solutions.

TNF-α (a product of R & D Systems, 10 μg/ml solution) was diluted withRPMI-1640 (containing 10% FCS) to prepare a 6 ng/ml TNF-α solution.Human aorta endothelial cells (HAEC) and human umbilical veinendothelial cells (HUVEC) were separately cultivated in a 96-wellculture plate, and when the cells became confluent, the medium in eachwell was removed. Then, 50 μl of each of the above-prepared testcompound solutions was added to the wells. To positive control wells andnegative control wells were added 50 μl of the medium and 100 μl of themedium, respectively. The plate was incubated at 37° C. for 30 minutes.50 μl of the TNF-α solution prepared above was added to all the wellsother than the negative control wells, and the plate was incubated at37° for 24 hours. The medium in each well was removed, and 100 μl ofparaformaldehyde (2% in PBS) was added to each well. Fixation wasconducted at room temperature for 10 minutes. After washing with aphysiological saline solution 6 times, a blocking solution (0.1% BSA(bovine serum albumin)/PBS) was added to each well. The plate wasincubated at room temperature for 1 hour. The blocking solution wasremoved. 100 μl of a primary antibody solution (the antibody diluted1,000-fold with 0.1% BSA/PBS) was added, and a reaction mixture wasincubated at 4° C. for 18 hours. After washing with a physiologicalsaline solution 5 times. 100 μl of a secondary antibody solution (theantibody diluted 1,000-fold with 0.1% BSA/PBS) was added, and a reactionmixture was incubated at room temperature for 2 hours. After washingwith a physiological saline solution 5 times, 100 μl of aperoxidase-labelled avidin solution (a product of DAKO Co., diluted1,000-fold with 0.1% BSA/PBS) was added. The reaction mixture wasincubated at room temperature for 1 hour. After washing with aphysiological saline solution 5 times, 100 μl of an OPD(o-phenylenediamine dihydrochloride) substrate solution was added andcolor development was allowed to take place at 37° C. Absorbancymeasurement at 492/692 nm was conducted, and a test compoundconcentration of 50% appearance inhibition of ICAM-1 or VCAM-1, i.e.IC₅₀ was determined.

As the test compound, the compound of Example 2 was used. The primaryantibody and the secondary antibody were as follows.

-   -   Primary antibody:        -   Mouse anti-human ICAM-1 (a product of Becton, Dickinson &            Co.)        -   Mouse anti-human VCAM-1 (a product of Becton, Dickinson &            Co.)        -   Secondary antibody:        -   Rabbit anti-human immunoglobulin (a product of DAKO Co.)

The results are shown in Table 14. TABLE 14 Human aorta Human umbilicalendothelial vein endothelial cells (μM) cells (μM) ICAM-1 40% inhibition25 at 100 μM VCAM-1 15 30% inhibition at 100 μMPharmacological Test 3 (TNF-α Production-Inhibiting Action)

A test compound was dissolved in 0.1 M sodium hydroxide. Thereto wasadded a 9-fold volume of PBS (a Dulbecco formula, a product of TakaraCo.) to prepare a 1 mM test compound solution. The solution was dilutedwith 0.1 M sodium hydroxide/PBS (1:9) to prepare 0.1 mM, 0.01 mM, 1 μM,0.1 μM and 0.01 μM test compound solutions.

A 50 μg/ml lipopolysaccharide (LPS) solution was prepared usingRPMI-1640 (containing 10% FCS). A 24-well culture plate was used. 1.35ml of RPMI-1640 (containing 10% FCS) was added to LPS-unstimulatedcontrol wells, and 1.32 ml of RPMI-1640 (containing 10% FCS) was addedto LPS-stimulated control wells. To the other wells were added 1.17 mlof RPMI-1640 (containing 10% FCS) and 0.15 ml of each diluted testcompound solution prepared above. To all the wells was added 0.15 ml ofwhole human blood, and the wells were incubated at 37° C. for 30minutes. Lastly, to all the wells other than the LPS-unstimulatedcontrol wells, was added 0.03 ml of the above-prepared LPS solution, andall the wells were incubated at 37° C. for 24 hours. Low-speedcentrifugation was conducted, and the supernatant in each well wascollected and measured for TNF-α concentration, by the use of acommercial ELISA kit. A test compound concentration of 50% TNF-αproduction inhibition, i.e. IC₅₀ was determined. The results are shownin Table 15. TABLE 15 Test compound IC₅₀ (μM) Compound of Example 2 10Compound of Example 3 6.4 Compound of Example 4 36.0 Compound of Example5 40.0 Compound of Example 6 33.5 Compound of Example 7 7.4 Compound ofExample 8 3.4 Compound of Example 9 0.7 Compound of Example 10 4.0Compound of Example 11 19 Compound of Example 12 5.7 Compound of Example13 7.5 Compound of Example 14 0.47 Compound of Example 15 2.3 Compoundof Example 16 2.7 Compound of Example 17 2.0 Compound of Example 18 2.3Compound of Example 19 0.88 Compound of Example 20 4.7Pharmacological Test 4 (IL-1 Production-Inhibiting Action)

An IL-1 production was measured in the same manner as in PharmacologicalTest 3, and a test compound concentration of 50% production inhibition,i.e. IC₅₀ was determined. When the test compound was the compound ofExample 2, the IC₅₀ was 80 μM.

Pharmacological Test 5 (IL-6 Production-Inhibiting Action)

An IL-6 amount produced was measured in the same manner as inPharmacological Test 3, and a test compound concentration of 50%production inhibition, i.e. IC₅₀ was determined. When the test compoundwas the compound of Example 2, the IC₅₀ was 100 μM or higher.

Pharmacological Test 6 (IFN-γ Production-Inhibiting Action)

A test compound was dissolved in 0.1 M sodium hydroxide. Thereto wasadded a 9-fold volume of PBS (a Dulbecco formula, a product of TakaraCo.) to prepare a 1 mM test compound solution. The solution was dilutedwith 0.1 M sodium hydroxide/PBS (1:9) to prepare 0.1 mM, 0.01 mM, 1 μM,0.1 μM and 0.01 μM test compound solutions.

A 50 mg/ml concanavalin A (Con A, a product of Seikagaku Co.) solutionwas prepared using RPMI-1640 (containing 10% FCS). A 24-well cultureplate was used. 1.35 ml of RPMI-1640 (containing 10% FCS) was added toCon A-unstimulated control wells, and 1.32 ml of RPMI-1640 (containing10% FCS) was added to Con A-stimulated control wells. To the other wellswere added 1.17 ml of RPMI-1640 (containing 10% FCS) and 0.15 ml of eachdiluted test compound solution prepared above. To all the wells wasadded 0.15 ml of whole human blood, and the wells were incubated at 37°C. for 30 minutes. Lastly, to all the wells other than the ConA-unstimulated control wells, was added 0.03 ml of the above-preparedCon A solution, and all the wells were incubated at 37° C. for 48 hours.Low-speed centrifugation was conducted, and the supernatant in each wellwas collected and measured for IFN-γ concentration, by the use of acommercial ELISA kit. A test compound concentration of 50% productioninhibition, i.e. IC₅₀ was determined. When the test compound was thecompound of Example 2, the IC₅₀ was 5 μM.

Preparation Example 1

Compound of Example 1 150 g Avicel (trade name for 40 g microcrystallinecellulose, a product of Asahi Chemical Industry Co., Ltd.) Corn starch30 g Magnesium stearate 2 g Hydroxypropylmethylcellulose 10 gPolyethylene glycol-6000 3 g Castor oil 40 g Ethanol 40 g

The present active ingredient compound, Avicel, corn starch andmagnesium stearate were mixed together and ground, and the mixture wasshaped into tablets by using a conventional pounder (R 10 mm) for sugarcoating. The tablets were coated with a film-coating agent consisting ofhydroxypropylmethylcellulose, polyethylene glycol-6000, castor oil andethanol, to prepare film-coated tablets.

Preparation Example 2

Compound of Example 2 150 g Citric acid 1.0 g Lactose 33.5 g Dicalciumphosphate 70.0 g Pluronic F-68 30.0 g Sodium lauryl sulfate 15.0 gPolyvinyl pyrrolidone 15.0 g Polyethylene glycol (Carbowax 1500) 4.5Polyethylene glycol (Carbowax 6000) 45.0 g Corn starch 30.0 g Dry sodiumlauryl sulfate 3.0 g Dry magnesium stearate 3.0 g Ethanol A requiredamount

The present active ingredient compound, citric acid, lactose, dicalciumphosphate, Pluronic F-68 and sodium lauryl sulfate were mixed together.

The mixture was sieved through a No. 60 screen. The sieved mixture waswet-granulated with an ethanol solution containing polyvinylpyrrolidone, Carbowax 1500 and Carbowax 6000. When necessary, ethanolwas added to convert the mixture into a paste-like mass. Corn starch wasadded, and mixing operation was conducted until uniform particles wereformed. The particles were passed through a No. 10 screen, then placedin a tray, and dried in an oven at 100° C. for 12-14 hours. The driedparticles were sieved through a No. 16 screen. Next, dry sodium laurylsulfate and magnesium stearate were added to the sieved particles. Themixture was compressed into core tablets of desired shape by using atablet machine.

The core tablets were treated with a varnish, and then talc was sprayedthereon for prevention of moisture absorption. On the surfaces of theresulting core tablets, an undercoat layer was formed. Varnish coatingwas made on the undercoat layer sufficient times so as to make thetablets suitable for internal use. Further, undercoat layer formationand smooth coating were conducted to make the coated tablets completelyround and smooth. Color coating was conducted until the tablet surfacescame to have a desired color. After drying, the coated tablets werepolished to obtain tablets of uniform gloss.

Preparation Example 3

Compound of Example 2 5 g Polyethylene glycol (mol. wt.: 4000) 0.3 gSodium chloride 0.9 g Polyoxyethylene sorbitan monooleate 0.4 g Sodiummetabisulfite 0.1 g Methylparaben 0.18 g Propylparaben 0.02 g Distilledwater for injection 10.0 ml

Parabens, sodium metabisulfite and sodium chloride were dissolved indistilled water of about half the above volume at 80° C. with stirring.The resulting solution was cooled to 40° C. In the solution weredissolved the present active ingredient compound, polyethylene glycoland polyoxyethylene sorbitan monooleate. To the resulting solution wasadded the remainder of distilled water to obtain a final volume. Thethus-obtained solution was sterilized by passing through an appropriatefilter paper, to prepare an injection.

1-18. (canceled)
 19. A method for inhibiting adhesion of neutrophils tovascular endothelial cells by administering to a patient suffering fromdiseases caused by acceleration of adhesion of neutrophils to vascularendothelial cells, wherein the disease is selected from the groupconsisting of endotoxin shock, ARDS, burn, asthma, viral myocarditis inacute phase, idiopathetic dilated cardiomyopathy, shift from systemicinflammatory response syndrome (SIRS) toward organ failure, multipleorgan failure, Crohn disease, hyper-γ-globulinemia, systemic lupuserythematosus (SLE), multiple sclerosis, metastasis of cancer,monoclonal B cell abnormal disease, polyclonal B cell abnormal disease,atrial myxoma, Castleman syndrome, primary glomerulonephritis, mesangialproliferative glomerulonephritis, cancerous cachexia, Lennander'slymphoma, psoriasis, atopic dermatitis, Kaposi's sarcoma appearing inAIDS, postmenopausal osteroporosity, sepsis and hepatitis an agent forinhibiting adhesion of neutrophils to vascular endothelial cellscomprising, as the active ingredient, a thiazole compound or apharmaceutically acceptable salt thereof of the formula:

wherein R¹ is a phenyl group which may have one or more lower alkoxygroups as a substituent on the phenyl ring; and R² is a heterocyclicring residue having 1-2 hetero atoms selected from the group consistingof nitrogen atom, oxygen atom and sulfur atom, which heterocyclic ringresidue may have, as a substituent on the heterocyclic ring, from 1 to 3groups selected from the group consisting of carboxyl group and loweralkoxy group.
 20. The method for inhibiting adhesion of neutrophils tovascular endothelial cells according to claim 19, wherein the activeingredient is6-[2-(3,4-diethoxyphenyl)thiazole-4-yl]pyridine-2-carboxylic acid. 21.The method for inhibiting adhesion of neutrophils to vascularendothelial cells according to claim 19, wherein the disease is selectedfrom the group consisting of endotoxin shock, ARDS, asthma, shift fromsystemic inflammatory response syndrome (SIRS) toward organ failure,multiple organ failure, Crohn disease, systemic lupus erythematosus(SLE), multiple sclerosis, cancerous cachexia, psoriasis, atopicdermatitis, sepsis and hepatitis.
 22. The method for inhibiting adhesionof neutrophils to vascular endothelial cells according to claim 21,wherein the active ingredient is6-[2-(3,4-diethoxyphenyl)thiazole-4-yl]pyridine-2-carboxylic acid.